PSCA CRISPR Activation Plasmid (h): sc-402050-ACT

Prostate stem cell antigen (PSCA) is a glycosylphosphatidylinositol (GPI)-anchored cell surface protein in the Ly6/uPAR family that contributes to epithelial differentiation, adhesion, and intercellular signaling. In human tissues, PSCA expression is enriched in prostate and other epithelial compartments and can modulate membrane microdomain organization and receptor-associated signaling at the cell surface. Dysregulated PSCA expression has been reported across multiple epithelial malignancies, where it is used as a molecular marker to interrogate tumor cell state, lineage programs, and microenvironmental interactions. PSCA is therefore relevant for studies of cell identity, membrane-proximal signaling, and transcriptional control in cancer biology.

PSCA CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSCA expression without altering the underlying DNA sequence.

PSCA CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSCA locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the PSCA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSCA expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSCA locus and enabling the study of PSCA-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSCA pathway restoration in tumor cells with silenced or reduced PSCA expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.